Klenke Ulrike, Taylor-Burds Carol
Cellular and Developmental Neurobiology Section, NIH/NINDS, Bethesda, Maryland, USA.
Curr Protoc Neurosci. 2012 Apr;Chapter 3:Unit 3.25.1-16. doi: 10.1002/0471142301.ns0325s59.
Primary cultures obtained from embryonic nasal placodes can maintain olfactory neurons, olfactory ensheathing cells, and large numbers of gonadotropin releasing hormone-1 (GnRH) neurons. Depending on the age of the starting material, one can examine cell interactions important for placode formation or neuronal migration and axonal outgrowth. When generated at E11.5 in mouse, neuronal migration and axon outgrowth away from the main tissue mass occurs. This area of the explant, the periphery, is only a few cells thick. This characteristic offers the opportunity to image single cells and axons and allows pharmacological and molecular manipulations as well as physiological recordings to be performed. Here, we describe a system for culturing nasal explants used in our laboratory. This model system provides a method for obtaining physiological cellular responses with post hoc immunohistochemistry, or gene expression studies, on cells arising from the nasal placode.
从胚胎鼻基板获得的原代培养物可以维持嗅觉神经元、嗅鞘细胞以及大量促性腺激素释放激素-1(GnRH)神经元。根据起始材料的年龄,可以研究对基板形成、神经元迁移和轴突生长重要的细胞相互作用。在小鼠胚胎第11.5天产生时,神经元会从主要组织块迁移并长出轴突。外植体的这个区域,即周边,只有几层细胞厚。这一特性为对单个细胞和轴突进行成像提供了机会,并且允许进行药理学和分子操作以及生理记录。在这里,我们描述了一种用于培养我们实验室中使用的鼻外植体的系统。这个模型系统提供了一种方法,可通过对源自鼻基板的细胞进行事后免疫组织化学或基因表达研究来获得生理细胞反应。